This application relates to the field of antibody fragments derivatized with polymers, and in particular to the use of such derivatization to increase the circulation half-lives of antibody fragment-polymer conjugates. This application also relates to the field of inflammatory diseases and asthma, and in particular to anti-IL-8 antibody treatment of inflammatory diseases and asthmatic diseases. This application further relates to humanized anti-interleukin-8 (IL-8) antibodies and to high affinity variants of such antibodies.
Modification of proteins with polyethylene glycol (xe2x80x9cPEGylationxe2x80x9d) has the potential to increase residence time and reduce immunogenicity in vivo. For example, Knauf et al., J. Biol. Chem., 263: 15064-15070 (1988) reported a study of the pharmacodynamic behavior in rats of various polyoxylated glycerol and polyethylene glycol modified species of interleukin-2. Despite the known advantage of PEGylation, PEGylated proteins have not been widely exploited for clinical applications. In the case of antibody fragments, PEGylation has not been shown to extend serum half-life to useful levels. Delgado et al., Br. J. Cancer, 73: 175-182 (1996), Kitamura et al., Cancer Res., 51: 4310-4315 (1991), Kitamura et al., Biochem. Biophys. Res. Comm., 171: 1387-1394 (1990), and Pedley et al., Br. J. Cancer, 70: 1126-1130 (1994) reported studies characterizing blood clearance and tissue uptake of certain anti-tumor antigen antibodies J., 2: A1479 (1995) reported that low molecular weight (5 or 10 kD) PEG attached to a sulfhydryl group in the hinge region of a Fabxe2x80x2 fragment reduced clearance compared to the parental Fabxe2x80x2 molecule.
Interleukin-8 (IL-8) is neutrophil chemotactic peptide secreted by a variety of cells in response to inflammatory mediators (for a review see Hebert et al. Cancer Investigation 11(6):743 (1993)). IL-8 can play an important role in the pathogenesis of inflammatory disorders, such as adult respiratory distress syndrome (ARDS), septic shock, and multiple organ failure. Immune therapy for such inflammatory disorders can include treatment of an affected patient with anti-IL-8 antibodies.
Sticherling et al. (J. Immunol. 143:1628 (1989)) disclose the production and characterization of four monoclonal antibodies against IL-8. WO 92/04372, published Mar. 19, 1992, discloses polyclonal antibodies which react with the receptor-interacting site of IL-8 and peptide analogs of IL-8, along with the use of such antibodies to prevent an inflammatory response in patients. St. John et al. (Chest 103:932 (1993)) review immune therapy for ARDS, septic shock, and multiple organ failure, including the potential therapeutic use of anti-IL-8 antibodies. Sekido et al. (Nature 365:654 (1993)) disclose the prevention of lung reperfusion injury in rabbits by a monoclonal antibody against IL-8. Mulligan et al. (J. Immunol. 150:5585 (1993)), disclose protective effects of a murine monoclonal antibody to human IL-8 in inflammatory lung injury in rats.
WO 95/23865 (International Application No. PCT/US95/02589 published Sep. 8, 1995) demonstrates that anti-IL-8 monoclonal antibodies can be used therapeutically in the treatment of other inflammatory disorders, such as bacterial pneumonias and inflammatory bowel disease.
Anti-IL-8 antibodies are additionally useful as reagents for assaying IL-8. For example, Sticherling et al. (Arch. Dermatol. Res. 284:82 (1992)), disclose the use of anti-IL-8 monoclonal antibodies as reagents in immunohistochemical studies. Ko et al. (J. Immunol. Methods 149:227 (1992)) disclose the use of anti-IL-8 monoclonal antibodies as reagents in an enzyme-linked immunoabsorbent assay (ELISA) for IL-8.
One aspect of the invention is a method of treating an inflammatory disorder in a mammal comprising administering to the mammal an effective amount of a conjugate consisting essentially of one or more antibody fragments covalently attached to one or more nonproteinaceous polymer molecules, wherein at least one antibody fragment comprises an antigen binding site that binds to human interleukin-8 (IL-8), and wherein the apparent size of the conjugate is at least about 500 kD.
Another aspect of the invention is a method of treating an asthmatic disorder in a mammal comprising administering to the mammal an effective amount of a conjugate consisting essentially of one or more antibody fragments covalently attached to one or more nonproteinaceous polymer molecules, wherein at least one antibody fragment comprises an antigen binding site that binds to human interleukin-8 (IL-8), and wherein the apparent size of the conjugate is at least about 500 kD.